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Sains Malaysiana 53(9)(2024):
http://doi.org/10.17576/jsm-2024-5309-20
Mechanism of Damnacanthal Induced Apoptosis in CEM-SS Cell Line
(Mekanisme Apoptosis Teraruh Damnacanthal dalam Titisan Sel CEM-SS)
BANULATA
GOPALSAMY, SAIFUL YAZAN LATIFAH* & HISYAM ABDUL HAMID
Department
of Biomedical Science, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang,
Selangor, Malaysia
Diserahkan: 9 Januari 2024/Diterima: 5 Ogos 2024
Leukaemia,
is cancer of organs that is responsible to produce blood specifically the
lymphatic system and bone marrow. Due to the harsh effects of currently used
cancer drugs, damnacanthal, an anthraquinone obtained from the roots of Morinda elliptica is tested as a potential anticancer agent.
This study reports on the participation of the p53, Bcl-2 and Bax in the apoptosis induced by of damnacanthal,
on T-lymphoblastic leukaemia (CEM-SS) cell. Cell viability and morphology was
tested with trypan blue assay, flow cytometry analysis detected the apoptotic
activity of damnacanthal, caspase colorimetric
protease assay tested the Caspase 2, 3, 6, 8, and 9’s involvement and
Enzyme-linked Immunosorbent Assay (ELISA) was carried out to quantify the Human
p53, Bcl-2, and Bax expression levels. Damnacanthal exhibited cytotoxicity at doses 10 and 30
µg/mL after 72 h of incubation. This study reports that damnacanthal arrested the cell at G2/M phase and initiates the apoptotic activity in the
cells treated with 30 µg/mL of damnacanthal for 72 h
through caspase 2 and 6 activation and not caspases 3, 8, and 9. Furthermore,
this anthraquinone induces apoptosis via p53-independent pathway. Damnacanthal also lowered Bcl-2 and increased Bax activity in CEM-SS cell lines. These anticancer
properties of damnacanthal makes it a potential agent
to treat T-lymphoblastic leukaemia.
Keywords: Anticancer;
apoptosis; CEM-SS; damnacanthal
Abstrak
Leukemia adalah kanser bagi organ yang bertanggung jawab
menghasilkan darah, terutamanya sistem limfa dan sum sum tulang. Disebabkan
kesan yang buruk oleh ubat kanser yang sedia ada, damnacanthal, salah satu
antrakuinon yang diperoleh daripada akar Morinda elliptica telah diuji
sebagai agen anti kanser yang berpotensi. Penyelidikan ini melaporkan
penglibatan p53, Bcl-2 dan Bax dalam apoptosis aruhan
damnacanthal, ke atas sel T-limfoblastik leukemia (CEM-SS). Kemandirian sel dan
morfologi telah diuji dengan ujian tripan biru, analisis sitometri aliran
mengesan aktiviti apoptosis damnacanthal, ujian protease kolorimetrik caspase
menguji penglibatan Caspase 2, 3, 6, 8 dan 9 dan Ujian Imunosorben Berkaitan
Enzim (ELISA) telah dijalankan untuk mengukur tahap pengekspresan p53, Bcl-2
dan Bax manusia. Damnacanthal menunjukkan sitotoksisiti pada dos 10 dan 30
µg/mL selepas 72 jam pengeraman. Kajian ini melaporkan bahawa damnacanthal
menahan sel pada fasa G2/M dan memulakan aktiviti apoptosis dalam sel yang
dirawat dengan 30 µg/mL damnacanthal selama 72 jam melalui pengaktifan caspase
2 dan 6 dan bukan caspases 3, 8 dan 9. Tambahan pula, antrakuinon ini mendorong
apoptosis melalui laluan bebas p53. Damnacanthal juga menurunkan Bcl-2 dan
meningkatkan aktiviti Bax dalam sel CEM-SS. Sifat antikanser damnacanthal ini
menjadikannya agen berpotensi untuk merawat leukemia T-limfoblastik.
Kata kunci: Antikanser;
apoptosis; CEM-SS; damnacanthal
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*Pengarang untuk surat-menyurat; email: latifahsy@upm.edu.my
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